ProjectSummary/Abstract:HPVisanecessarycauseof10%ofallcancersofwomenglobally,including 99% of cervical cancers. Rates of cervical cancer vary markedly by geographic region and level of economic development,with>85%occurringindevelopingcountries.SuccessfulprophylacticvaccinationagainsthrHPV typescanpreventdysplasiaandthuscervicalcancer.GlobalestimatesofHPVvaccinationdeliveryandcervical screeningbyregionandincomelevelshowvirtuallynosignificantdeliverytomanypoorerpopulationsofwomen worldwide.Thereare13hrHPVtypesplusagroupof11possiblycarcinogenictypes.LicensedHPVvaccines alltargetHPV16andHPV18,andonealsotargetsthenextfivemostcommontypesincervicalcancer.Since theydonottargetallcancer-associatedHPVtypes,screeningmustbemaintainedeveninvaccinatedwomen, adding to medical costs. In addition, the stringent cold-chain requirements of HPV vaccines provide a serious impedimenttotheirdeliveryindevelopingcountries.OuroverallgoalistodevelopanaffordableHPVvaccine that both extends the breadth of protection to all cancer-associated HPV types, and is stable at ambient temperature.Hereweaddressthisgoalwithtwoinnovations:1)displayinganL2-basedprotectiveepitope(RG1) thatisconservedforHPVsinauniquepositiononthesurfaceofHPV16L1VLPs,producingasingleantigen, broadlyprotectiveHPVvaccinetechnology,RG1-VLP,and2)theRG1-VLPvaccineisformulatedonalumand embeddedinglassyorganicmatricesformedbyadjustinglyophilizationandformulationparametersinorderto control ice crystal nucleation rates, glass transition temperatures, and other material properties and thereby protectagainstdegradationduringprocessing,shippingandstorage. HYPOTHESIS1:ColdchainstoragepropertiesareasignificantbarriertothedistributionofcurrentHPVvaccines, andthedevelopmentofapowderedRG1-VLPvaccineincludingadjuvantthatisthermostablewilladdressthis need. Specific Aim 1: Develop a GLP freeze-dry protocol for a powder formulation of RG1-VLPs in alum and study its in vitro temperature stability, and in murine models its immunogenicity and protective efficacy in comparisontoGardasil9. HYPOTHESIS2:RG1-VLPvaccinationissafeandwelltolerated.SpecificAim2:ToperformaDoseEscalation PhaseITrialoftheSafetyandImmunogenicityofthermostableRG1-VLPin36healthyfemalevolunteerswith theinclusionofacontrolGardasil9arminthestudy. HYPOTHESIS3:RG1-VLPvaccinationofhealthywomeninducesbroadlyprotectiveserumantibody.Specific Aim3:ToanalyzethelevelsofprotectiveantibodiesintheserumofpatientsfromthephaseIstudyinducedby RG1-VLPvaccinationorGardasil9.Wewillutilizethepassivetransferassaytomeasureprotectiveresponses, as well as L2 and L1 VLP ELISA and in vitro neutralization assays to quantify antigen-specific neutralizing antibodytitersinserum.